Determination of Marker Compound of Artocarpus champeden Spreng. Stembark Extract And Validation of The Analysis Method Using HPLC Repository - UNAIR REPOSITORY

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Ethnobotanical, Proximate, and Phytochemical Studies of Areco vestiorio Giseke
(Pinang Yaki)
Herny E.l. Simbala And Trina

E.

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Tallei

optimization of Topical Patch containing of Kaempferia Oil
(Koemperia golongol

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Lucia Hend riati

Determination

of

Marker Compound of Artocorpus champeden

Spreng.

2L

Stembark Extract And Validation of The Analysis Method Using HPLc

Ni Putu

Ariantari, Lidya Tumewu, Agriana Rosmalina Hidayati, Aty

Widyawaruyanti, Achmad Fuad Hafid

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lsolation of Oligomerstilbenoids From Stem Bark of Vatico flovolirens Sloot

29

Endemic of South East Sulawesi
Siti Hadijah Sabarwati, Nohong, Euis H. Hakim, Yana M. Syah, Sutriadi S, Zaeni A

Antimicrobial And Anticancer Activity of Curcusone B lsolated From Stem Bark
of !otropha curcas
l. Sahidin, Ardiansyah, M. Taher, S. Arsad,S.J.A. lchwan, J. Latip, B.M. Yamin, and

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S.N. Eaharum

Screening of Medicinal Plants of South Sumatera

Against Organism From Skin lnfection
Salni, H anifa Marisa, Harmida

for Antibacterial Activity

Long Term Effect of Ethanolic Extract of Fenugreek Seeds

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(Trigonella foenum-gruecum L.) on White Rat Liver Function
Kurnia Agustini, Sriningsih, Lestari Rahayu

Analysis of Plasma Malondialdehide (Mda) Concentration By Giving Shallot,s
Extortion Waler lA!lium oscdlonicum L.) cn Hypercholesteroiemic Mice
lsmawati, Enikarmila Asni, M.Yulis Hamidy

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Chemical Constituents from Shorea faguetiano(Dipterocarpaceae)

Rohaiza Saat, Laily B. Din, Wan Yaacob Wan Ahmad

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DETERMINATION OF N{ARKER COMPOTIND OF
ARTqCARPUS CTIAMPEDEN SPRENG. STEMBARK EXTRACT
AND VALIDATION OF THE ANALYSIS METHOD USING
HPLC
Ni Putu Ariantarir, Lidya Tumervu2. Agriana Rosnralina IIidayati2.
Aty Widyarvaruyantir. Achmad Fuad Hafidl.
'Phar,r,ucy Department, Faculty of Mathematics and Natural Sciences, Udayrnr Urrivcrsiry; Jalan
Kampus Bukit Jimbaran Badung-Bali Indonesia 80364; Phone: 081 337 017 28? OR 0361-

703837; e-mail: ari dedhika @.vahoo.com
2Postgraduated
Student, Faculty of Pharmacy, Airlangga University, Surabaya
lPhytochemistry anC Pharmacognosy Department, Faculty of Pharmacy, Airlangga University,
Surabaya, Indonesia

.\bstract : Artocarpus clnnpedel SPRENG. (Moraceae), commonly known as "cempedak", has
been traditionally used for malarial remedies. Several studies also reported in vitro and in vivo
antimalarial activity of this plant. In order to devcloped A. champedetr srembark as anrimalarial
phytopharmaceutical product, marker approach was chosen for quality control purpose of the
exhact as mw material.
The aims of this study are to determine marker compound of A. clompeden stembark
extract and develop an analysis method for quantification of marker compound in ethanol extract
of A. champeden SPRENG. stcmbark using HPLC.
Isolation of marker isolate was caried out by column chromatography and prepantive
TLc rechniques. Based on fIV, IR, 'H NMR, "c NMR, 2D NMR (CosY, HMBC and HMQC)
and MS spectra and comparison with other refcr:nces, confirmed that marker isolatc was identical
with known prcnylatcd chalcone, Morachalconc A. Thc HPLC profile showed that marker
compound has time retcntion shown as Mean t SD (RSD) of 12.547 minutes 1 0.39 (3.08%).
Validation of analysis method for quantification of marker in ethanol extract was conducted. The

analyte were chromatographed on C8 250x4.6 mm column, methanol-water as mobile phase
(65:35 v/v) by isocratic elution at flow rate of I m[-/minute, column temperature of 30'C, stop
time of analysis at 20 minutes and detection wavclength at 385 nm. The method was validated in
terms of selectivity, spccificity, linearity, accuracy and precision. The calibration curvc was linear
within a high range concentration of 25 - 50 pgm.L-l and low range conccnuation of 3-5 - X
pgml--r. The accuracy and intraday precision wcrc found to be well within acccptable limits. The
marker content in ethanol extract ofA. champeden SPRENG. stembark was shown as M?all I SD
(RSD) of 0.33617o I 0.03 (8.45%).

Keywords: A. clumpeden SPRENG., antimalarial activity, ethanol extmct, markcr compound,
morachalcone A, HPLC analysis.

INTRODUCTION
Artocarplts champeden SPRENG. belongs to Moraceae, commonly known as
"cempedak", is widely distributed in Indonesia and has been traditionally used for
maiarial remedies [Hakim et aI.,2006]. Previous studies reported that several prenylated
flavonoids isolated from A. champeden SPRENG. has potent antimalarial activity in vitro
against P. faLciparunr strains 3D7 [3] and in vivo against P. berghei strains ANKA
(unpublished). This indicates lhat A. chantpeden is prospective to be developed as
phytomedicine product for antimalarial.

The pharmaceutical requirements for a herbal product destined for a multi-target
therapy are very comp)ex [Kroll and Cordes, 2006]. The constituents of medicinal herbs
can vary greatly as a result of genetic factors, climate, soil quality and other external

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Coderence on fuledicina{rPhnts - SuroSaya,Indoneia 21-21

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factors ISchulz et al.,2004]. Therefore, despite the use of authenticated botanical voucher
specimens to help assure proper identity, modern concepts and methods relating to the

quality (i.e., chemical consistency)

of

herbal materials and products pertain to

phytochemical markers and fingerprint analyses. These markers are the threads that tie
together production and quality control [Reif et al., 2004]. Ideally, chemical markers
should be unique components that contribute to the therapeutic effects of a herbai
medicine [Li et al., 2008].
The standardized extract should have consistent constituent in order to ensure the
consistency of quality, safety and efficacy of the product. The marker approach to ensure
consistency is based upon the assumption that the content of other constituents rvill vary
in proportion to the marker compound, that if each batch contains the same standardize
amount of marker, the content of other constituents will also be relatively consistent
ILazarowych and Pekos, 19981 McCutcheon, 2002].
In order to develop A. chompeden as antimalarial phytomedicine product with
multi-component approach, it is needed to standardize A. chantpeden extract as raw
material. Therefore, the study to obtain standardized extract of A. chantpeden SPRENG.
stembark as raw material of antimalarial phytomedicine product using active marker
compound was ccnducted.

The preliminary study showed that 80% ethanol extract of A.

champeden

stembark was active extract that inhibit parasit growth both in vitro against Plasnodiunt
falcipanim strains 3D7 and in vivo againsi Plasnroditun berghei strains ANKA. Then,
from this ethanol extract has been identified an isolate belongs to chalcone group, based
on IJV spectra, as characteristic orange spot, detected in extract, fraction and subfraction
using TlC-Densitometer and HPLC. This isolate also showerf, in vitro antimalarial
aciivity against P. falciparum strains 3D7 with an ICjg value of0.69 pglml-, therefore this
isoiate can be used as marker candidate from 807o ethanol extract of A. champeden
stembark.

The present study aims to determine and isolate marker compound from 807o
ethanol extract of A. champeden stembark, then develop an analysis method for
quantiication of marker content in ethanol extract of A. champeden stembark. Therefore,
in the beginning we isolate and identify the stucture of an isolate that we used as marker
candidate for ethanol extract of A. champeden stembark. This study consist of several
steps, are begun with isolation of marker compound using conventional column
chromatography and preparative TLC techniques, followed by identification of isolate
based on uv-Vis, IR, 'H NMR, '3c Mt4R, 2D NMR (cosY, HMBC and HMQC) and
MS spectra. Further step is development of analysis method for quantification of marker
content in ethanol extract of A. champeden stembark using HPLC that conducted by
validation of an analysis method.

MATERIALS AND METHODS
Plant material
The stembark of A rtocarpus champeden SPRENG. was collected from Bogor, West Java,
Indonesia, in June 2007 and 2008. A voucher specimen rvas identified and deposited at
the Herbarium Bogoriense, Bogor Botanical Garden, Bogor, Indonesia.
Chemicals and Reagents
All reagents were analytical grade, otherwise reagents for HPLC analysis. Methanol for
HPLC analysis was purchased from Mallinckodt, inc. (USA). Samples for HPLC were
filtered through a 0.45 pm membrane filter (Millipore, Bedford, MA, USA). Reference
standard, marker compound Morachalcone A, was isolated from the stembark of
Artocarpus champeden SPRENG. In our laboratory; and mass spectrometrv, NMR

spectrometry and infrared spectrometry were applied

characteristics.

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rproceefrng of latemationot Conference on *lefrcinaftPhats - gurafiala,Iaf,otesia 21-21

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General Experimental Techniques and Instruments
Silica gel was used for columr chromatography (Merck, 0.063-0.200 mm) and silica gel
for preparative thin layer chromatography (Merck). Spots on plates were detected under
UV light (L 254 and 366 nm) and by spraying with l0% HzSOc in water followed by
gentle heating. IR spectrum obtained on a Shimadzu spectrometer IR Prestige-2l type.
The NMR experiments (both lD and 2D) were obtained on a Jeol spectrometer ECA 500
type operating at 500 MHz. HPLC analysis was performed with Hewlett Packard Agilent
I 100 series, an Agilent [ 100 series Degasser Gl322A, a Rheo-dyne 7725 injection valve
with a 20-pL loop, Agilent I 100 series Quartenary Pump Gl3l lA, Agilent I100 series

Column Comparlement Gl3l6A, Agilent 1100 series diodearay detector (DAD)

Gl3l5A.

Compounds were separated on a 250x4.6 mm Varian Microsorb

MV

100-5 C8

column.

Extraction and Isolation
One kilogram of Artocarpus champeden SPRENG. stembark was extracted with
807o ethanol at 60"C in rotavapor, yielded 74,6368 g of crude extract. This extract was
applied to ODS column chromatography, using methanol-water (4:l v/v)-methanol.,rethanol-acetonitril (l:l v/v) and acetonitril as eluent, resulting in l0 major fractions
(fraction l-10). Fraction 4 (686.9 mg) was applied to silica gel column and eluated with
CHCI3 followed by increasing polarity of CHCITMeOH I -l\Vo by gradient elution,
yielded 9 nrajor subfraction. Further separation of subfraction 4.8 (47.1 mg) was
conducted with several steps of Preparative TLC techniques using silica RP-CIS as
stationary phase with MeOH-H2O (7:3 vlv) mixtures as mobile phase yielded active
subfraction 4.8.8 ( 18.7 mg). This was then continued using silica as stationary phase with
CHCI:-MeOH (9.5:0.5 v/vi mixtures as mobile phase yielded active subfraction 4.8.8.7
(12.2 mg). Purification of this subfraction was conducted by reverse-phase PTLC with
MeOH-HzO (4: I v/v) mixtures as mobile phase resulting in active marker isolate,
Morachalcone A (7.2 mg).
Morachalcone A: orange powder; W (MeOH) i,*. nm 250,316 and 385; IR
(KBr) cnrr: 3,456, 1,697. rH and '3C I.IMR data are given in Table l.
Antimalaria! activity Testing
The antimalarial activity of fractions and the isolated compound was determined by the
procedure described by Budimulya et al. (1997\. In brief, each fractions and compound
was separately dissolved in DMSO (10-2 mol L-'; and kept at -zO'C until used. The
maladal parasite P. falciparum 3D7 clone was propagated in a 24-well culture plate in the
presence of a wide range of concentration of each fractions or compound. The growth of
the parasite was monitored by making a blood smear fixed with MeOH and stained with
Giemsa stain. The antimalarial activity of each fractions or compound was expressed as
an IC50 value, defined as the concentration of the compound causing 507o inhibition of
parasite growth relative to an untreated control.

HPLC Conditions
HPLC analysis was conducted with methanol-water (65:35 v/v) as mobile phase by
isocratic elution, C8 250x4.6 mm Varian Microsorb MV 100-5 column at flow rate of I
ml/minute, column temperature of 30"C, stop time of analysis at 20 minutes and detection
vt)avelength set at 385 nm.

Preparation of standard solution
The standard stock solution ( I mg/ml) was prepared by dissolving 5 mg of standard to
mL of methanol. The stock solution was stored at 4'C.

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Sample preparation
Ten mg of ethanol extract of A. chantpeden SPRENG stembark was accurately weighed
and dissolved in methanol using vortex for 5 minutes. Sample solution then filtered
through a 0.45 pm syringe membrane filter.

Calibration
The standard stock solutions of marker substance was diluted by methanol to give serial
concentrations are: at 25i 30; 35;40;45 dan 50 pgmL-r thigh range concentration) and at
25 20: lO;7; 5.6 dan 3.5 pgml--' (low range concentration). After filtering through a
0.45 pm mernbrane filter, 20 pL of each concentration was injected into the HPLC
column for lnalysis. Linear regression model was used to describe the relationship
between peak area and concentration of standard solution.

Mcthod validation
lvlethod validation for quantification of marker conrent in ethanol extract of A.
chanpeden stembark using HPLC was carried out in terms of selectivity, specificity,
linearity, limit of detection, limit of quantitation, accuracy and precision.
Selcctivity and Specificity
Selectivity and specificity was checked by using an extract of Artocarpus chantpeden
SPRENG. stembark and marker compound Morachalcone A and optimizing separation
and detection. The purity of the peaks was checked by acquisition of spectra ()" = 200400 nm) by use of the DAD detector. Spectra were acquired at the upslope, apex, and
downslope of each peak, computer normalized, and superimposed. Peaks were considered
pure when there was coincidence between the three spectra (match factor >950.000).
Selectiviy study resulting in methanol-water (65:35 v/v) as mobile phase by isocratic
elution, C8 250x4.6 mm Varian Microsorb MV l0O-5 column at flow rate of I
rnlJminute, column temperature of 30'C, stop time of analysis at 20 minutes and
deteclion wavelenSth set at 385 nm.

Limits of Detection and Limit of Quantification
Limits of detection and quantification were determined as the amounts for which the
calculated signal-to-noise ratios were approximately

3:

I

and 10:1, respectively.

Precission
Standard stock solution was diluted in methanol for intra-day test (injecting certain
concentration of standard six times within 24 hours). The standard deviation (SD) and
relative standard deviation (RSD) were calculated.

Accuracy
Accuracy strrdy was carried out by spiking certain concentration of marker to sample
Recovery rate (7o) was calculated.

RESULT AND DISCUSSION
The dried stem bark of A. champeden was extracted with 807o ethanol. In a
preliminary test of in vitro antimalarial activity against P. falciparum 3D7 clone of this
ethanol extract showed significant inhibition (unpublished). Marker isolation from
ethanol extract of A. champeden stembark was done by bioactivity guided fractination.
The ethanol extract was fractionated with open column chromatography using ODS as
stationary phase and metanol-water as mobile phase, resulting in l0 major fractions and
fraction 4 rvas proved to be active against P. falcipan{m strains 3D7 in vitro. Fraction 4
was applied to silica gel column and eluated with CHCI3 followed by increasing polarity
of CHCI3-MeOH l-l\Ea by gradient elution, yielded active subfraction 4.8 with an IC56
value of 0.03 gg/n,L. Further separation oi subfraction 4.8 with several steps of PTLC

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techniques using silica RP-Cl8 as stationary phase with MeOHIHzO mixtures as mobile
phase yielded active subfraction 4.8.8 (ICso value of 0.10 pglml-), then silica as stationary

with CHCI3-MeOH mixtures as mobile phase yielded active subfraction 4.8.8.7
value of 0.39 pglml). Purihcation of this subfraction by reverse-phase PILC with

phase
(ICso

MeOH-H2O mixtures as mobile phase resulting in active marker isolate.
r3C
}',IMR,2D NMR
Identification of isolate based on W-Vis, IR, 'H NMR,
(COSY, HMBC and HMQC) spectra and comparison with other references. Marker
compound was isolated as orange powder. The [FV-Vis absorptions at 250,316 and 385
nm were suggestive of a c[alcone skeleton [Mabry, et al., 1970]. The IR spectrum of
morachalcone A contained- absorption bands at 3,456 crrr and 1,697 cm-r corresponding
rH Nl"iR
to hydroxyl and cdrbonyl groups, respectively [Silverstein et al., l99l]. The
spectrum contained charactheristic signals ascribable to an isoprenyl group (6H 1.65,
1.17, 5.22, 3.34). Two proton signals at EH 7.73 (lH, dd, J=15.25 Hz) and 8.1 (lH, dd,
J=15.9 Hz) form an AB system, the large coupling constant indicating the trans geometry
of a double bond. The 'H-I.IMR spectrum (table l) also indicated signals for ortho
coupled aromatic protons in ring A (6 6.36 (ZH, m) dan 6 7.52 (lH, dd, J= 8.55,3.7 Hz)
and two aromatic protons in ring B 6 6.43 (lH, q, J=8.55, 17.75 Hz) dan 6 7.76 (lH, d,
J=".55 Hz). The presence of proton signals f,t 6 13.93 and 6 14.48 indicated that the
hydroxyl groups are loocated at C-2 ard C-4 in ring A and C-2' and C-4' in ring B. The
''C-Mv{R spectrum contained signals from 20 carbon atoms including that of a ketone
carbonyl carbon at 6c 194.25 (table l).
On the basis of HMQC and HMBC spectral analysis, all protons and carbon
signals were fully assigned and the positions of the substituents on tbe aromatic rings
were determinb. The HMBC correlations for H-1"/C-2', C-3' and C-3" confirmed that

the 3,3{imethylallyl group is located at C-3'. Structue determination for marker
compound also conducted based on comparison with their spectroscopic data from
literature values [Abegaz et al., 1998; Monache et al., 1995]. Thus, the structure of
marker compound was deduced as known prenylated chalcone, Morachalcone A.
Morachalcone A was previously isolated from callus culture of Maclura pomifera
(Moraceae) [Monache et al., 1995] and also from methanol extract of A- champeden
stembark (unpublished)
Tabel 1. The rH I.IMR and 13C NMR 500
C

o
D

,H

7;13 dd, (15.25 Hz't
8.1 dd ( 15.9 Hz)

C=O

3

4
6

,,C

t17.93
142.15
194,25
t 14.68

1

5'

data for marker com ound

6.43 q (8.55, l7 .75 Hz)
7.76 d (8.55 Hz\

163.50
116.62
165.32
108:26
1,30.47

115.74
160.85
103.69

I

2
3

6.36 m

5

6

6.36 m
7.52 dd (8.55, 3.7 Hz)

cHz

3.3,1

22.60

CH=

5.22

123.70

E-Me

1.65

s

Z-Me

1.77

s

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t62.7't
r09.23
132.56

13t.59
26.09
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intcrnal standard. Only signals that significanr for the comparison are reported

1l

H

5

OH

'('--,/
o

Fig.

I

Chemical structure of actiye marker compound isolated from
Artocarpus c hanpeden SPRENG.

The marker compound revealed inhibitory activity against P. falciparum strain
3D7 rvith an IC5o value of 0.18 pglml. The inhibitory activity of this compound showed
that the isolated compound was an active marker. The result of this study showed that
active marker compound, lvlorachalcone A, can be used to
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Figure 1. Chromatogram obtained from ethanol extract of A. champeden slembark (Eet) and
marker standard morachalcone A (Std)

standardize ethanol extract

of A.

champeden stembark

as

antimalarial

phytomedicine product. Morachalcone A, obtained from this study was then used as
reference standard for analysis of rnarker content in ethanol extract of A- champeden
stembark using HPLC. According to Reif [2004], marker compound can be isolated in
c',vn laboratory if it is not comercially available and used as reference standard with
defined identification, purity and content.
The development of analysis method for quantification of marker content in
ethanol extract of A. champeden stembark using HPLC wds conducted by validation of an

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Woceeding oJ InterndtionaI Conference on 5llef,icinaIrPfan* - SuraSala,Iaf,oretia 21-21

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2010

analysis method, Validation of the method was done in terms of selectivity, specificity,
linearity, accuracy, precision, limit of detection and limit of quantitation.
The chromatographic condition was optimized through comparison of different
solvents and solvent ratio. Compared with other solvents, the mixture of methanol and
water (65:35 v/v) as mobile phase by isocratic elution, C8 250x4.6 mm Varian
Microsorb MV 100-5 column at flow rate of I mljminute, columr temperature of 30"C,
stop time of analysis at 20 ninutes and detection wavelength set at 385 nm showed the
best separation and minimize peak tailing.
Under this condition, the chromatogram profile of ethanol extract (Figure l)
showed that peak at time retention shown rs Mean ! SD (RSD) of 13.001 minutes a 0.37
Q-87qo), has resolution of 1.42, plate number of 3524, peak width at half height of
0.5200, symmetry factor of 0.82 and selectivity factor of l. I l. The purity factor of this
peak was 997.034, indicated that targeted peak fall within acceptable purity. The identity
of this peak against marker spectrum was shown as match factor value of 981.261
(>950.000), indicated that targeted peak analyzed in ethanol extract was marker
compound that has been detected in ethanol extract of A. champeden stembark. Linearity
study shorved that analysis within the high range concentration of 50 - 25 pgml'' and low
range concentratio,. of 3.5 - 25 pgml--r resulting in respcnse proportional against
concentration with correlation coefficient G) of 0.9989 and 0.9997, V^e value of l.26Vo
and 2.llVo respectively, indicated linear conelation. Limit of detection was 1.57 pgml--'
and limit of quantitaion was 4.71 pgml.-r, The compound analysis were cbtained with the
recovery rate shown as Mean ! SD (RSD) of 102.5991Vo ! 5.62 (5.48Eo), with intra-day
for instrument precision shown as Mean ! SD (RSD) of 31.0668 + 1.88 (6.06 7o) and
method precision shown as RSD of 8.457o. Based on parameter values obtained from
validation study, the analysis method can ie assumed to be valid for quantification of
marker content in ethanol extract of A. champeden stembark using HPLC. Amount of
Morachalcone A as marker in ethanol extract of /'r. champeden stembark shown as Mean
t SD (RSD) was 0.33617o + 0.03 (8.45Vo).

CONCLUSION
Results of this study showed that prenylated chalcone, Morachalcone A, can be used as
marker for ethanol extract of Anocarpus champeden SPRENG. stembark. HPLC method

also has been developed for quantitation of Morachalcone

A in

ethanol extract of A.

champeden SPRENG. stembark. Results indicate that the present method was valid in
controlling the quality of ethanol extract of A. champeden SPRENG. stembark. The
quantitative analysis showed that the content of marker Morachalcone A in ethanol
extract ofA. champeden was shown asMean!SD (RSD) of 0.336170 10.03 (8.4570).

ACKNOWLEDGMENTS
This work was supported by Grant for Competition Research (no. 5641103.UPG12007)
from Directorate General of Higher Education, The Ministry of National Education,
Republic of I-ndonesia.

R.EFERENCES
Abegaz BM, Ngandjui BT, Dongo E, Tamboue H., 1998, Prenylated Chalcones and
Flavones from The Leaves of Dorstenia kameruniana. Phytochemistry, 49(4),

I147-l150
Budimulya A.S., Syafruddin, Tapchaisri P., Wilariat P., Marzuki S., 1997, The sensitivity

of

Plasmodium protein synthesis

Biochem Parasitol, 84,

137-

l4l

to prokaryotic ribosomal inhibitors, Mol

2l

Qroceefrng of Intend,tionaf Conference on X'tediciru[Qhn* - SuraSaya,Intonesia 21-21

1 q 2010

Hakim 8.H., Achmad S.A., Juliawaty L.D., Makmur L., Syah Y.M., Aimi N., Kitajima
M., Takayama H., Ghisalberti E.,2006, Prenylated Flavonoids and Related
Compounds ofTt Indonesian Artocarpus (Moraceae), J Nat Med, 60, l6l-184
Kroll U., Cordes C.,2006, Pharmaceutical prerequisites for a multi-target therapy,
Phytomedicine, 13, 12-19
Lazarowych N.J., Pekos P.,1998. Use of Fingerprinting and Marker Compounds for
Identification and Standardization of Botanical Drugs: Strategies for Applying
Pharmaceutical HPLC Analysis to Herbal Product, Drug Information Journal, 32,
197-512

Li

S., Han Q., Qiao C., Song J., Chen, C.L., Xu H., 2008, Chemical Markers for The
Quality Control of Herbal Medicine: An Overview, Chinese Medicine, 3(7), l -16
Mabry T.J., Markham K.R., Thomas M.B., 1970, The Systematic Identification of
Flavonoids. Springer-Verlag, New York-Heidelberg-Berlin, pp 227 -241
Mccutcheon A.R.,2002, An Exploration of Current Issues in Botanical Quality: A
Discussion Paper. Canada, |{atural Health Products, pp l-[ I
Monache G.D., De Rosa M.C., Scurria R., Vitali A., Cuteri A., Monacelli B., Pasqua G.,
Botta B., 1995, Comparison Between Metabolite Productions in Cell Culture and
in Whole Plant of Maclura po,...fera, Phytochemistry 39(3), 575-580
Reif K., Sievers H., Steffen J.P., 2004, The Role of Chemical Reference Standards as
Analytical Tools in the Quality Asessment of Botanical Materials - An European
Perspective, HerbalGram, 63. 3843
Schulz V., liansel R., Blumenthal M., Tyler V.E., 2004, Rational Phytotherapy:
Reference Guide for Physicians and Pharmaciss, Fifth Edition, Springer-Verlag,
Hiedelberg. pp t- l0
Silverstein -R.M., Bassler G.C., Monill T.C., 1991, Spectrometric Identification of
Organic Compounds, John Wiley & Sons Inc, Singapore, pp 3-10;91-97
Widya.waruyanti A., Subehan, Kalauni S.K., Awale S., Nindatu M., Zaini N.C.,
Syafruddin D., Asih P.B.S., Tezuka Y., Kadota S., 2007, New Prenylated
Flavones frcm Artocdrpus champeden SPRENG. and Their Antimalarial Activity
In Vitro, J Nat Med, 61, 410413

28

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UI\IV T'I(SI TAS AII(LANGGA
FAKULTAS FARMASI
Dharn'"wsngsa DaleE Surab.F J0286 Tclp.

Websitc : htto://qqu.lLunair.ac.id

03 1-5Ol37lO

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SURAT TUGAS
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Sehubungan dengan adanya Internatlonal Conference On Medicinal
Plants The Future of Medicinal Plants : Form Plant to Medicine pada
tanggal 2L - 22 luli 2010 di Fakultas Farmasi Universitas Widya
Mandala, dengan lnl Wakil Dekan II Fakultas Farmasi Universitas
Airlangga menugaskan :

- Dr. Aty lArldyawaruyanti,MSl
- Dr. Achmad Fuad H.MS

NrP. 19620426 199002 2 001
NIP. 19521212 198103 1 009

Untuk menghadlri acara tersebut.

Demikian Surat Penugasan lni untuk dilaksanakan dengan baik dan

penuh tanggung Jawab.

Surabaya, 20

luli 2010

kan

kil Dekan

.".i

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a

,

A*-

.H. Umi Athiyah,MS
NrP. 19s60407 198103 2 001

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Deutsrher Akademkher Austausddienst

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6erman Atademic Exchanqe 5ervice

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This is to certifythat

Achmad

Fr^rael

has participated

tlafirl

in

38o Meeting of Nqtional Working Group on Indonesian Medicinal Plants

INTERNATIONAL CONFERENCE ON MEDICINAI PLANTS
Surabaya,

Jily 2l

-

22, 2OlO

as

ORAL PRESENTER

The Nadonal Wortlng Group
on Indoneclaa Medtclnal Plants

[lE
Chairman

M.

la.l".

nat. Elisabedr C.Wlc

Chairman